Title:AP Lab #2: Enzyme CatalystsIntroduction:

Enzymes are catalytic proteins, meaning they speed up – but do not create – chemicalreactions, without being used up or altered permanently in the process. Although variousenzymes use different methods, all accomplish catalysis by lowering the free energy of activation – activation energy – for the reaction, thus allowing it to occur more easily.Enzymes employ a variety of methods for performing catalysis. Some provide a microenvironment within the active site where some of the side chains are H+ or OH- donors or receivers. Other enzymes work by bringing together substrates that would not normally meetoutside the enzyme, or orienting them in a manner in which they would otherwise not occur. Stillother enzymes stress the bonds of substrate molecules in order to make them easier to break;some take this a step further by actually forming temporary covalent bonds with the substratemolecules. Regardless of how it is done, all enzyme catalyzed reactions are reversible and willturn around when necessary.As a result of four levels of organization, an enzyme has a very specific shape, which iscalled its conformation. Even more specific is the active site of the enzyme, where the actualcatalysis occurs. The specific molecule or closely related molecules on which an enzymefunctions is known as its substrate. Shape plays such an important role in enzymatic catalysis,that often even isomers of the substrate will be rejected. Once the substrate enters the active site,it may begin a process known as induced fit in which the enzyme perfectly conforms to themolecule to allow for more efficient catalysis.Enzymes have specific environmental conditions at which they will function best. As aresult, changes in environment can severely impact enzyme catalysis in both negative and positive ways. Each enzyme has specific ranges at which it optimally functions; in general,increasing the temperature will help the reaction along, until the point at which the proteindegrades and denatures – or falls apart into its lower level structures. Denatured proteins willoften return to their original state, after the removal of the denaturing agent, except when theyare degraded multiple levels. The rate of reaction through catalysis can also be increased byincreasing the concentration of either the enzyme or the reactants; enzyme if all the active sitesare full or the substrate if the active sites are not all full.

Hypothesis:

I predict that the reaction will begin rather quickly with the addition of the enzyme, but will slow as the H

is converted to H

O and O

I also believe that, given the amount of time the reactions are to be run for, not all of the H

will react, thus leaving some to react withthe KMnO

even after the full 360 seconds.

Materials:

Beaker, H

, KMnO

, water (boiling and unboiled), potato or liver, paper,syringe or pipette, burette, mixing stick, and a stopwatch.

Procedure:

Add 10 ml of H

to each of the labeled beakers for the set time they will be measuring.

Add 1 ml of H

0 to the first beaker.

Allow the reaction to occur for the set time limit labeled on the specified beaker.

Once the time limit is fulfilled, stop the reaction by adding 10 ml of H

to the beaker.

Repeat this procedure for all beakers with their specified time limit and mix well.

Gather a 5 ml sample from on of the beakers and transfer it to a new glass beaker.

Record initial burette reading containing KMnO

Continue to add KMnO

until a faint brown color can be seen.

Record final burette reading.

10)

Calculate the ml of KMnO

used to reach the end of the titration.

11)

Repeat this procedure for the rest of the other time-labeled beakers.

Data:Exercise 2A:

1. a) Potassium Iodide.1. b) Hydrogen Peroxide1. c) Water and Oxygen1. d) The bubbles forming2. The temperature denatured the protein making the enzyme useless.3. The H

reacts with the liver by fizzing up. If the liver were boiled, there would be minimalto no reactions occurring.

Exercise 2B:Base Line Calculation

Final Reading of Burette 1.2 mLInitial Reading of Burette5.0 mLBase Line3.8 mL

Exercise 2C:Uncatalyzed Hydrogen Peroxide Decomposition

Final Reading of Burette3.2 mLInitial Reading of Burette10.0 mLAmount of KMnO

6.8 mL

Exercise 2D:Table 2.1:KMnO

(mL)Time (seconds)

10 s30 s60 s90 s120 s180 sa) Base Line4.3 ml4.3 ml4.3 ml4.3 ml4.3 ml4.3 ml b) Final Reading1.8 ml1.8 ml2.0 ml2.0 ml2.4 ml3.0 mlc) Initial Reading5.0 ml5.0 ml5.0 ml5.0 ml5.0 ml5.0 mld) Amount of KMnO

Consumed3.2 ml3.2 ml3.0 ml3.0 ml2.6 ml2.0 mle) Amount of H

Used1.1 ml1.1 ml1.3 ml1.3 ml1.7 ml2.3 ml

Graph 2.1:Analysis of Results:

Time Intervals (seconds)

0 to 1010 to 3030 to 6060 to 9090 to 120120 to 180

Rates*

0.11 ml0.00 ml0.00667 ml0.00 ml0.01333 ml0.01 ml2. At the beginning. When the hydrogen peroxide and catalase are first mixed, the substrateconcentration is high and the molecules meet active sites often and with consistency.3. Near the end. As the hydrogen peroxide is decomposed into H

O and O

, there is less of thesubstrate to bind with the active site, thus slowing the reaction. Had there still been sufficientH

for the active sites, the reaction would not have slowed.4. Sulfuric acid acts to inhibit the function of catalase because it denatures the enzyme whichremoves its catalytic abilities. As we know, enzymes have very specific and their structures arespecially developed (conformed) to fit the molecule with which it reacts (substrate). When the